What would the consequence be if you used only a forward primer or a reverse primer for a Sanger dideoxy-sequencing reaction and NOT both the forward and reverse primers in one reaction?

A. You would have 2 overlapping sequences from each DNA strand and not be able to decipher which bases followed which, thus not being able to decipher any sequence.

B. You would generate a sequence read only half the length of a read using both primers.

C. You always use 2 primers when doing PCR, whether you are sequencing DNA or not! You won’t have any sequence to read if you don’t!

D. You would be doing it correctly.

2. What step must you do before sequencing or PCR-amplifying RNA? (with the exception of nanopore sequencing technology)

A. You just need to isolate clean, pure RNA. (that’s it)

B. You need to create cDNA from RNA using Taq polymerase

C. You need to create cDNA from RNA using reverse transcriptase

D. You need to isolate DNA as well as RNA from your sample.

3. Which statement about exome sequencing is false?

A. It is a way to eliminate sequencing about 99% of the genome.

B. It is justified because most of the harmful SNPs (mutations) are in coding regions.

c. It is sequencing only the introns of a genome and not the exons.

d. You can use exome capture techniques in order to eliminate the non-coding regions from the sequencing pool.

e. None. All of the above are true.

4. Which statement about reporters is false?

A. Lab-worthy GFP emits a green fluorescence when it binds and cleaves its substrate when illuminated with UV light.

B. Luciferase enzymes use luciferin substrates to yield a light (photon) signal; they are often used to measure promoter activity.

C. Beta-galactosidase is an enzyme encoded by the lacZ gene and cleaves galactosides like ONPG and X-gal to yield yellow and blue colors, respectively.

D. Alkaline phosphatase is an enzyme encoded by the phoA gene that cleaves ONPP to yield a yellow color and XP to yield blue.

E. None. All of the above are true.

5. Sanger’s chain termination sequencing is the method that has been used most often in the last 40 years. Which statement about it is true?

A. The reactions incorporate dideoxy nucleotide triphosphates (ddNTPs) throughout the growing nucleotide chain during extension by the polymerase.

B. The reactions terminate extension with a radioactively or fluorescently labeled dNTP.

C. All 4 G, A, T, and C reactions can be run in the same tube as long as each ddNTP has a different “colored” tag (a tag that can be read at a different wavelength)

D. The reactions are run in miniaturized, massively parallel flow cells to get maximum read lengths.

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